Wavenumber-dependent dynamic light scattering optical coherence tomography measurements of collective and self-diffusion.

We demonstrate wavenumber-dependent DLS-OCT measurements of collective and self-diffusion coefficients in concentrated silica suspensions across a broad q-range, utilizing a custom home-built OCT system. Depending on the sample polydispersity, either the collective or self-diffusion is measured. The measured collective-diffusion coefficient shows excellent agreement with hard-sphere theory and serves as an effective tool for accurately determining particle sizes. We employ the decoupling approximation for simultaneously measuring collective and self-diffusion coefficients, even in sufficiently monodisperse suspensions, using a high-speed Thorlabs OCT system. This enables particle size and volume fraction determination without the necessity of wavenumber-dependent measurements. We derive a relationship between the particle number-based polydispersity index and the ratio of self and collective mode amplitudes in the autocorrelation function and utilize it to measure the particle number-based polydispersity index. Notably, the polydispersity determined in this manner demonstrates improved sensitivity to smaller particle sizes compared to the standard intensity-based DLS cumulant analysis performed on dilute samples.

Lensless single-shot dual-wavelength digital holography for industrial metrology.

We demonstrate lensless single-shot dual-wavelength digital holography for high-speed 3D imaging in industrial inspection. Single-shot measurement is realized by combining off-axis digital holography and spatial frequency multiplexing of the two wavelengths on the detector. The system has 9.1 µm lateral resolution and a 50 µm unambiguous depth range. We determine the theoretical accuracy of off-axis dual-wavelength phase reconstruction for the case of shot-noise-limited detection. Experimental results show good agreement with the proposed model. The system is applied to industrial metrology of calibrated test samples and chip manufacturing.

Experiment and theory of the complex refractive index of dense colloidal media.

The complex refractive index is analyzed by measuring its scattering attenuation µ s, group index n g, and group velocity dispersion (GVD) for 100 nm diameter silica nanoparticles dispersed in water. The experiments were performed for wavelengths between 410 nm and 930 nm. The experimental results were compared with different mixing models for the complex refractive index of colloidal suspensions. The group index linearly scaled with the volume fraction both in experiment and for all tested models. It was found that the GVD has a nonlinear dependence on volume fraction in agreement with the coupled dipole model of Parola et al. [J. Chem. Phys.141, 124902 (2014)JCPSA60021-960610.1063/1.4895961] The scattering attenuation is in good agreement with both the coupled dipole model and the low frequency quasi-crystalline approximation [J. Electromagn. Waves Appl.2, 757 (1988)JEWAE50920-507110.1163/156939388X00053] that take particle correlations into account. With an iterative fitting procedure of all the data based on both the coupled dipole model and the quasi-crystalline approximation, the refractive index, porosity, and size of the nanoparticles were determined. We determined that the coupled dipole model is in best agreement with the data.

Measuring optical properties of clear and turbid media with broadband spectral interferometry.

The group index, n g, group velocity dispersion (GVD), and scattering attenuation coefficient, µ s, were measured for dilutions of glycerol, ethanol, and Intralipid 20% with water. Experiments were performed with a supercontinuum laser based Mach-Zehnder spectroscopic interferometry setup for wavelengths between 400 and 930 nm. All optical properties could be retrieved from a single calibrated measurement of the interference spectrum. Scattering attenuation was determined from the envelope of the interference. The group index and GVD were retrieved from the unwrapped spectral phase. It was found that the group indices of glycerol and ethanol dilutions are in accordance with the Lorentz-Lorenz mixing formula. The scattering attenuation matches well to a semi-empirical model based on the Twerksy effective packing fraction.

Computational 3D resolution enhancement for optical coherence tomography with a narrowband visible light source.

Phase-preserving spectral estimation optical coherence tomography (SE-OCT) enables combining axial resolution improvement with computational depth of field (DOF) extension. We show that the combination of SE-OCT with interferometric synthetic aperture microscopy (ISAM) and computational adaptive optics (CAO) results in high 3D resolution over a large depth range for an OCT system with a narrow bandwidth visible light super-luminescent diode (SLD). SE-OCT results in up to five times axial resolution improvement from 8 µm to 1.5 µm. The combination with ISAM gives a sub-micron lateral resolution over a 400 µm axial range, which is at least 16 times the conventional depth of field. CAO can be successfully applied after SE and ISAM and removes residual aberrations, resulting in high quality images. The results show that phase-preserving SE-OCT is sufficiently accurate for coherent post-processing, enabling the use of cost-effective SLDs in the visible light range for high spatial resolution OCT.

Sub-diffusion flow velocimetry with number fluctuation optical coherence tomography.

We have implemented number fluctuation dynamic light scattering optical coherence tomography (OCT) for measuring extremely slow, sub-diffusion flows of dilute particle suspensions using the second-order autocovariance function. Our method has a lower minimum measurable velocity than conventional correlation-based OCT or phase-resolved Doppler OCT, as the velocity estimation is not affected by the particle diffusion. Similar to non-dilute correlation-based OCT, our technique works for any Doppler angle. With our analysis we can quantitatively determine the concentration of particles under flow. Finally, we demonstrate 2D sub-diffusion flow imaging with a scanning OCT system at high rate by performing number fluctuation correlation analysis on subsequent B-scans.

Scanning dynamic light scattering optical coherence tomography for measurement of high omnidirectional flow velocities.

We show scanning dynamic light scattering optical coherence tomography (OCT) omnidirectional flow measurements. Our method improves the velocity measurement limit over conventional correlation-based or phase-resolved Doppler OCT by more than a factor of 2. Our technique is applicable without a-priori knowledge of the flow geometry as our method works both for non-zero Doppler angle and non-ideal scan alignment. In addition, the method improves the particle diffusion coefficient estimation for particles under flow.

Roadmap on Digital Holography-Based Quantitative Phase Imaging.

Quantitative Phase Imaging (QPI) provides unique means for the imaging of biological or technical microstructures, merging beneficial features identified with microscopy, interferometry, holography, and numerical computations. This roadmap article reviews several digital holography-based QPI approaches developed by prominent research groups. It also briefly discusses the present and future perspectives of 2D and 3D QPI research based on digital holographic microscopy, holographic tomography, and their applications.

Fast and accurate spectral-estimation axial super-resolution optical coherence tomography.

Spectral-estimation OCT (SE-OCT) is a computational method to enhance the axial resolution beyond the traditional bandwidth limit. However, it has not yet been used widely due to its high computational load, dependency on user-optimized parameters, and inaccuracy in intensity reconstruction. In this study, we implement SE-OCT using a fast implementation of the iterative adaptive approach (IAA). This non-parametric spectral estimation method is optimized for use on OCT data. Both in simulations and experiments we show an axial resolution improvement with a factor between 2 and 10 compared to standard discrete Fourier transform. Contrary to parametric methods, IAA gives consistent peak intensity and speckle statistics. Using a recursive and fast reconstruction scheme the computation time is brought to the sub-second level for a 2D scan. Our work shows that SE-OCT can be used for volumetric OCT imaging in a reasonable computation time, thus paving the way for wide-scale implementation of super-resolution OCT.

Attenuation coefficient estimation in Fourier-domain OCT of multi-layered phantoms.

Optical properties, such as the attenuation coefficients of multi-layer tissue samples, could be used as a biomarker for diagnosis and disease progression in clinical practice. In this paper, we present a method to estimate the attenuation coefficients in a multi-layer sample by fitting a single scattering model for the OCT signal to the recorded OCT signal. In addition, we employ numerical simulations to obtain the theoretically achievable precision and accuracy of the estimated parameters under various experimental conditions. Finally, the method is applied to two sets of measurements obtained from a multi-layer phantom by two experimental OCT systems: one with a large and one with a small Rayleigh length. Numerical and experimental results show an accurate estimation of the attenuation coefficients when using multiple B-scans.

Analysis of attenuation coefficient estimation in Fourier-domain OCT of semi-infinite media.

The attenuation coefficient (AC) is an optical property of tissue that can be estimated from optical coherence tomography (OCT) data. In this paper, we aim to estimate the AC accurately by compensating for the shape of the focused beam. For this, we propose a method to estimate the axial PSF model parameters and AC by fitting a model for an OCT signal in a homogenous sample to the recorded OCT signal. In addition, we employ numerical analysis to obtain the theoretical optimal precision of the estimated parameters for different experimental setups. Finally, the method is applied to OCT B-scans obtained from homogeneous samples. The numerical and experimental results show accurate estimations of the AC and the focus location when the focus is located inside the sample.

Quantification of plant morphology and leaf thickness with optical coherence tomography.

Optical coherence tomography (OCT) can be a valuable imaging tool for in vivo and label-free digital plant phenotyping. However, for imaging leaves, air-filled cavities limit the penetration depth and reduce the image quality. Moreover, up to now quantification of leaf morphology with OCT has been done in one-dimensional or two-dimensional images only, and has often been limited to relative measurements. In this paper, we demonstrate a significant increase in OCT imaging depth and image quality by infiltrating the leaf air spaces with water. In the obtained high-quality OCT images the top and bottom surface of the leaf are digitally segmented. Moreover, high-quality en face images of the leaf are obtained from numerically flattened leaves. Segmentation in three-dimensional OCT images is used to quantify the spatially resolved leaf thickness. Based on a segmented leaf image, the refractive index of an infiltrated leaf is measured to be 1.345±0.004, deviating only 1.2% from that of pure water. Using the refractive index and a correction for refraction effects at the air-leaf interface, we quantitatively mapped the leaf thickness. The results show that OCT is an efficient and promising technique for quantitative phenotyping on leaf and tissue level.

Multi-scale optical coherence tomography imaging and visualization of Vermeer's Girl with a Pearl Earring.

We demonstrate multi-scale multi-parameter optical coherence tomography (OCT) imaging and visualization of Johannes Vermeer's painting Girl with a Pearl Earring. Through automated acquisition, OCT image segmentation, and 3D volume stitching we realize OCT imaging at the scale of an entire painting. This makes it possible to image, with micrometer axial and lateral resolution, an entire painting over more than 5 orders of length scale. From the multi-scale OCT data we quantify multiple parameters in a fully automated way: the surface height, the scattering strength, and the combined glaze and varnish layer thickness. The multi-parameter OCT data of Girl with a Pearl Earring shows various features: Vermeer's brushstrokes, surface craquelure, paint losses, and restorations. Through an interactive visualization of the Girl, based on the OCT data and the optical properties of historical reconstructions of Vermeer's paint, we can virtually study the effect of the lighting condition, viewing angle, zoom level and presence/absence of glaze layer. The interactive visualization shows various new painting features. It demonstrates that the glaze layer structure and its optical properties were essential to Vermeer to create an extremely strong light to dark contrast between the figure and the background that gives the painting such an iconic aesthetic appeal.

Polarization contrast optical diffraction tomography.

We demonstrate large scale polarization contrast optical diffraction tomography (ODT). In cross-polarized sample arm detection configuration we determine, from the amplitude of the optical wavefield, a relative measure of the birefringence projection. In parallel-polarized sample arm detection configuration we image the conventional phase projection. For off-axis sample placement we observe for polarization contrast ODT, similar as for phase contrast ODT, a strongly reduced noise contribution. In the limit of small birefringence phase shift δ we demonstrate tomographic reconstruction of polarization contrast images into a full 3D image of an optically cleared zebrafish. The polarization contrast ODT reconstruction shows muscular zebrafish tissue, which cannot be visualized in conventional phase contrast ODT. Polarization contrast ODT images of the zebrafish show a much higher signal to noise ratio (SNR) than the corresponding phase contrast images, SNR=73 and SNR=15, respectively.

Large-scale high-sensitivity optical diffraction tomography of zebrafish.

In this work we demonstrate large-scale high-sensitivity optical diffraction tomography (ODT) of zebrafish. We make this possible by three improvements. First, we obtain a large field of view while still maintaining a high resolution by using a high magnification over numerical aperture ratio digital holography set-up. With the inclusion of phase shifting we operate close to the optimum magnification over numerical aperture ratio. Second, we decrease the noise in the reconstructed images by implementing off-axis sample placement and numerical focus tracking in combination with the acquisition of a large number of projections. Although both techniques lead to an increase in sensitivity independently, we show that combining them is necessary in order to make optimal use of the potential gain offered by each respective method and obtain a refractive index (RI) sensitivity of 8 ⋅ 10 - 5 . Third, we optimize the optical clearing procedure to prevent scattering and refraction to occur. We demonstrate our technique by imaging a zebrafish larva over 13 mm 3 field of view with 4 micrometer resolution. Finally, we demonstrate a clinical application of our technique by imaging an entire adult cryoinjured zebrafish heart.

Comparison of image reconstruction techniques for optical projection tomography.

We present a comparison of image reconstruction techniques for optical projection tomography. We compare conventional filtered back projection, sinogram filtering using the frequency-distance relationship (FDR), image deconvolution, and 2D point-spread-function-based iterative reconstruction. The latter three methods aim to remove the spatial blurring in the reconstructed image originating from the limited depth of field caused by the point spread function of the imaging system. The methods are compared based on simulated data, experimental optical projection tomography data of single fluorescent beads, and high-resolution optical projection tomography imaging of an entire zebrafish larva. We demonstrate that the FDR method performs poorly on data acquired with high numerical aperture optical imaging systems. We show that the deconvolution technique performs best on highly sparse data with low signal-to-noise ratio. The point-spread-function-based reconstruction method is superior for nonsparse objects and data of high signal-to-noise ratio.

Sub-millimeter depth-resolved digital holography.

We present sub-millimeter full-field depth from focus digital holography of surface topography of rough objects. For each pixel, the depth of the object is calculated from the variance of the intensity image over a set of reconstruction distances. First, we theoretically describe the axial resolution of this method and show that sub-millimeter resolution is feasible. Second, using a digital holography setup without magnifying optics or lateral scanning we experimentally demonstrate 100 µm axial resolution depth ranging and surface topography imaging. This is significantly better than what has previously been reported using digital holography and could make this technique useful for rapid large-area characterization of surface topography of objects.

Point spread function based image reconstruction in optical projection tomography.

As a result of the shallow depth of focus of the optical imaging system, the use of standard filtered back projection in optical projection tomography causes space-variant tangential blurring that increases with the distance to the rotation axis. We present a novel optical tomographic image reconstruction technique that incorporates the point spread function of the imaging lens in an iterative reconstruction. The technique is demonstrated using numerical simulations, tested on experimental optical projection tomography data of single fluorescent beads, and applied to high-resolution emission optical projection tomography imaging of an entire zebrafish larva. Compared to filtered back projection our results show greatly reduced radial and tangential blurring over the entire [Formula: see text] mm2 field of view, and a significantly improved signal to noise ratio.

Wavefront sensorless adaptive optics OCT with the DONE algorithm for in vivo human retinal imaging [Invited].

In this report, which is an international collaboration of OCT, adaptive optics, and control research, we demonstrate the Data-based Online Nonlinear Extremum-seeker (DONE) algorithm to guide the image based optimization for wavefront sensorless adaptive optics (WFSL-AO) OCT for in vivo human retinal imaging. The ocular aberrations were corrected using a multi-actuator adaptive lens after linearization of the hysteresis in the piezoelectric actuators. The DONE algorithm succeeded in drastically improving image quality and the OCT signal intensity, up to a factor seven, while achieving a computational time of 1 ms per iteration, making it applicable for many high speed applications. We demonstrate the correction of five aberrations using 70 iterations of the DONE algorithm performed over 2.8 s of continuous volumetric OCT acquisition. Data acquired from an imaging phantom and in vivo from human research volunteers are presented.

Image resolution and deconvolution in optical tomography.

We present a frequency domain analysis of the image resolution of optical tomography systems. The result of our analysis is a description of the spatially-variant resolution in optical tomographic image after reconstruction as a function of the properties of the imaging system geometry. We validate our model using optical projection tomography (OPT) measurements of fluorescent beads embedded in agarose gel. Our model correctly describes both the radial and tangential resolution of the measured images. In addition, we present a correction of the tomographic images for the spatially-varying resolution using a deconvolution algorithm. The resulting corrected tomographic reconstruction shows a homogeneous and isotropic pixel-limited resolution across the entire image. Our method is applied to OPT measurements of a zebrafish, showing improved resolution. Aside from allowing image correction and providing a resolution measure for OPT systems, our model provides a powerful tool for the design of optical tomographic systems.